A subset of human papillomavirus (HPV) types have been implicated in cervical cancer. The determination of the presence of the HPV types associated with cervical cancer may serve as adjunct to Pap smear screening of cervical samples. This analysis could reduce the incidence of invasive cervical cancer, while decreasing the human and economic costs associated with Pap smear screening and reflex histological analysis. HPV analysis could distinguish between transient infections that resolve without the development of high-grade cervical lesions, and the presence of rare cells that have been biologically altered by HPV, and are progressing from a localized intraepithelial lesion to invasive cancer. This project will develop a method using Rolling Circle Amplification to analyze individual cells within a cervical sample and establish the presence of HPV DNA and RNA sequences, and in Phase II, establish the relationship of other markers to the early stages of HPV-related cervical cancer. The majors goals of this project are to develop a means to detect rare HPV-transformed cells that are missed by conventional cytology and validate RCA protocols for molecular analyses of cells with respect to features of HPV gene expression that present a risk of neoplastic progression rather than a transient infection. PROPOSED COMMERCIAL APPLICATIONS: The successful completion of this project will provide two types of products. The first could serve as an adjunct Pap smears by providing an indication when a "second look" might reveal rare aberrant cervical cells missed by cytology. The ultimate product would enhance or replace the annual Pap smear by providing molecular information, predicting early cervical cancer lesions, that is not visible to a trained cytotechnologist or pathologist.